Revvity, 64PLCG1TPEH, HTRF Human and Mouse Total PLC-γ1 Detection Kit, 10,000 Assay Points

Overview
This HTRF cell-based assay conveniently and accurately detects Phospholipase C gamma 1 (PLCgamma 1) protein.
PLCgamma1 is a phosphoinositide-specific phospholipase that generates second messengers and plays a crucial role in signal transduction. Upon cell stimulation, PLCgamma 1 interacts with tyrosine kinase receptors and others, like T cell receptors, leading to the activation by phosphorylation at Ty783.
PLCgamma 1 is a potential target in the context of a number of diseases. For example, frequent mutations in the PLCG1 gene have been reported in angiosarcoma and T-cell lymphomas and its activation has also been implicated in the spreading of breast cancer.
How it works
Total-PLCg1 assay principle
The Total PLCg1 assay quantifies the expression level of PLCg1 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-PLCg1 assay uses two labeled antibodies, one coupled to a donor fluorophore and the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In the presence of PLCg1 in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein's expression under a no-wash assay format.

 

Total-PLCg1 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates to a 384-well low volume detection plate before the addition of Total-PLCg1 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total-PLCg1 one-plate assay protocol
Detection of Total PLCg1 with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4 Lysis Buffer 5
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-10,000 assay points